Efficient and wash-free labeling of membrane proteins using engineered Npu DnaE split-inteins

Protein Science : a Publication of the Protein Society
Euiyeon LeeYoungeun Kwon

Abstract

An efficient and wash-free method to conjugate a fluorescent tag to a target membrane protein is developed, using engineered Npu DnaE split-inteins. This approach allowed fast labeling while avoiding the strenuous synthesis of a long polypeptide. Two different modes of labeling, namely specific binding and covalent conjugation, are observed. The covalent labeling was monitored within 5 min, without background staining.

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Citations

Aug 26, 2020·Proceedings of the National Academy of Sciences of the United States of America·Vedud PurdeDmitri S Kudryashov
Oct 16, 2019·Proceedings of the National Academy of Sciences of the United States of America·Maniraj BhagawatiHenning D Mootz
Jan 12, 2021·Chembiochem : a European Journal of Chemical Biology·Philipp WolfOliver Seitz

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